Biochemical and electrophoretic analyses of saliva from the predatory reduviid species Rhynocoris marginatus (Fab.).

The saliva of Rhynocoris marginatus consists of amylase, invertase, trehalase, protease, acid phosphatase, alkaline phosphatase, phospholipase, lipase, trypsin, hyaluronidase, and esterase. All enzyme activities were significantly higher in the saliva of female R. marginatus when compared to the saliva of male individuals. The saliva was analyzed by tricine SDS/PAGE, sephadex column chromatography, FT-IR, and MALDI-TOF. The pH of the saliva was slightly alkali. The SDS/PAGE revealed a few proteins with molecular masses greater than 29.5 and 36.2 kDa for male and female predator saliva respectively. The FT-IR spectrum confirmed the acidic, proteinaceous, enzymatic, and aromatic nature of the saliva. The MALDI-TOF-MS revealed the presence of enzymes, proteins, peptides, and other biomolecules. The most prominent peptides were named as RmIT-1 (3.79 kDa), RmIT-2 (9.7 kDa), and RmIT-3 (10.94 kDa) (Rhynocoris marginatus Insect Toxin). Further studies are underway to isolate and identify these biomolecules.

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Enzymatic Digestion and Mass Spectroscopies of N-Linked Glycans in Lacquer Stellacyanin fromRhus vernicifera

International Journal of Polymer Science ◽

10.1155/2015/547907 ◽

2015 ◽

Vol 2015 ◽

pp. 1-9 ◽

Cited By ~ 1

Author(s):

Oyunjargal Tumurbaatar ◽

Takashi Yoshida

Keyword(s):

Enzymatic Digestion ◽

Acetone Powder ◽

Blue Color ◽

Sephadex Column ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Glycosylation Sites ◽

Pngase F ◽

C 50 ◽

Tof Ms

Lacquer stellacyanin was isolated and purified from lacquer acetone powder by continuous Sephadex column chromatographies using Sephadex C-50, DEAE A-50, and C-50 gels. The purified lacquer stellacyanin had a blue color with one major and three minor bands around 26 k Dain SDS PAGE. Trypsin- and chymotrypsin-treated lacquer stellacyanins were examined by LC/MS/MS to determine three N-glycosylation sites (N28, N60, and N102) and were further analyzed by MALDI TOF MS, indicating that the N-linked glycans were attached to the three asparagine (Asn) sites, respectively. In addition, after trypsin digestion and PNGase A and PNGase F treatments to cleave N-linked glycans from the Asn sites, it was found that lacquer stellacyanin had a xylose containing a biantennary N-linked glycan with core fucosylation consisting of 13 sugar residues (a complex type N-linked glycan) by MALDI TOF MS analysis. This is the first report on the structure of an N-linked glycan in lacquer stellacyanin.

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Electroelution of Intact Proteins from SDS-PAGE Gels and Their Subsequent MALDI-TOF MS Analysis

Plant Proteomics ◽

10.1385/1-59745-227-0:353 ◽

2006 ◽

pp. 353-364 ◽

Cited By ~ 1

Author(s):

Zhentian Lei ◽

Ajith Anand ◽

Kirankumar S. Mysore ◽

Lloyd W. Sumner

Keyword(s):

Maldi Tof Ms ◽

Intact Proteins ◽

Maldi Tof ◽

Ms Analysis ◽

Sds Page ◽

Tof Ms

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2155. Accelerated Confirmation of Porin Loss in Carbapenem-Resistant Enterobacterales: A MALDI-TOF Mass Spectrometry-Based Approach

Open Forum Infectious Diseases ◽

10.1093/ofid/ofz360.1835 ◽

2019 ◽

Vol 6 (Supplement_2) ◽

pp. S731-S731

Author(s):

Carlos Correa-Martinez ◽

Evgeny A Idelevich ◽

Karsten Becker

Keyword(s):

Mass Spectrometry ◽

Molecular Weight ◽

Gold Standard ◽

Sodium Dodecylsulfate ◽

Resistant Bacteria ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Carbapenem Resistant ◽

Sds Page ◽

Tof Ms

Abstract Background The accurate identification of carbapenem resistance mechanisms is decisive for the appropriate selection of antibiotic regimens. Numerous methods can detect carbapenemase-producing carbapenem-resistant bacteria (CPCR). However, non-CPCR (NCPCR) are routinely assumed to display porin loss as a diagnosis of exclusion. No further confirmatory tests are performed since the gold standard (sodium dodecylsulfate polyacrylamide gel electrophoresis, SDS–PAGE) is laborious and time consuming. We propose a test for rapid and easy detection of porin loss by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Methods Clinical meropenem-resistant Enterobacterales strains (10 CPCR, 10 NCPCR) and control strains recommended by EUCAST (5 carbapenemase-producing, one with porin loss, one-negative control) were analyzed. Membrane proteins were extracted by successive centrifugation of bacterial suspensions (McFarland 0.5) and addition of ethanol, formic acid and acetonitrile. MALDI-TOF MS of the protein extracts was performed on a 96-spot target (Bruker Daltonics, Germany). Peaks between 35 and 40 kDa were analyzed for the presence of porins and compared with the bands observed in the SDS–PAGE of the protein extracts. Results Within the molecular weight range of 35–40 kDa, the MALDI-TOF MS-based method revealed peaks in all CPCR isolates corresponding to those observed in the carbapenemase-producing control strains. In contrast, the control strain with porin loss as well as all CNCR isolates showed a lower quantity of peaks in this range. All peaks observed correlated with the bands observed in the SDS–PAGE of the protein extracts at the corresponding molecular weight (Figure 1). Conclusion Yielding results that reliably correspond to the current gold standard, we propose a method for accelerated detection of porin loss as an alternative to the diagnosis of exclusion usually made in routine settings. With a processing time of approximately 20 minutes, the method can be easily implemented in the clinical setting. Applying this MALDI-TOF MS-based approach, valuable information will be provided about a resistance mechanism that otherwise remains unexplained. Disclosures All authors: No reported disclosures.

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Washing-free electrochemical DNA detection using double-stranded probes and competitive hybridization reactionElectronic supplementary information (ESI) available: HPLC and MALDI-TOF MS data of the synthetic ferrocene-conjugated signaling DNA. Grazing angle FT-IR spectra. Sequences of PCR mimics. See http://www.rsc.org/suppdata/cc/b4/b402914c/

Chemical Communications ◽

10.1039/b402914c ◽

2004 ◽

pp. 1466 ◽

Cited By ~ 23

Author(s):

Kyuwon Kim ◽

Haesik Yang ◽

Se Ho Park ◽

Dae-Sik Lee ◽

Sung-Jin Kim ◽

...

Keyword(s):

Ir Spectra ◽

Dna Detection ◽

Grazing Angle ◽

Supplementary Information ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Ft Ir ◽

Competitive Hybridization ◽

Electrochemical Dna Detection ◽

Tof Ms

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A novel mechanism of “metal gel-shift” by histidine-rich Ni2+-binding Hpn protein from Helicobacter pylori strain SS1

10.1101/081331 ◽

2016 ◽

Author(s):

Rahul Mahadev Shelake ◽

Yuki Ito ◽

Junya Masumoto ◽

Eugene Hayato Morita ◽

Hidenori Hayashi

Keyword(s):

Helicobacter Pylori ◽

Metal Binding ◽

Sodium Dodecyl ◽

Polyacrylamide Gel ◽

Metal Species ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Sds Page ◽

Gel Shift ◽

Tof Ms

AbstractSodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) is a universally used method for determining approximate molecular weight (MW) in protein research. Migration of protein that does not correlate with formula MW, termed “gel shifting” appears to be common for histidine-rich proteins but not yet studied in detail. We investigated “gel shifting” in Ni2+-binding histidine-rich Hpn protein cloned from Helicobacter pylori strain SS1. Our data demonstrate two important factors determining “gel shifting” of Hpn, polyacrylamide-gel concentration and metal binding. Higher polyacrylamide-gel concentrations resulted in faster Hpn migration. Irrespective of polyacrylamide-gel concentration, preserved Hpn-Ni2+ complex migrated faster (3-4 kDa) than apo-Hpn, phenomenon termed “metal gel-shift” demonstrating an intimate link between Ni2+ binding and “gel shifting”. To examine this discrepancy, eluted samples from corresponding spots on SDS-gel were analyzed by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF-MS). The MW of all samples was the same (6945.66±0.34 Da) and identical to formula MW with or without added mass of Ni2+. MALDI-TOF-MS of Ni2+-treated Hpn revealed that monomer bound up to six Ni2+ ions non-cooperatively, and equilibrium between protein-metal species was reliant on Ni2+ availability. This corroborates with gradually increased heterogeneity of apo-Hpn band followed by compact "metal-gel shift" band on SDS-PAGE. In view of presented data metal-binding and “metal-gel shift” models are discussed.

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Isolation, Purification, and Identification of Coconut Protein through SDS-PAGE, HPLC, and MALDI-TOF/TOF-MS

Food Analytical Methods ◽

10.1007/s12161-020-01743-1 ◽

2020 ◽

Vol 13 (6) ◽

pp. 1246-1254

Author(s):

Yuan Lin ◽

Yan Wang ◽

Zherong Ji ◽

Xueyi Le

Keyword(s):

Maldi Tof ◽

Sds Page ◽

Coconut Protein ◽

Tof Ms

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Synthesis and Characterization of Copoly(Ether Sulfone)s with Different Percentages of Diphenolic Acid Units

Polymers ◽

10.3390/polym12081817 ◽

2020 ◽

Vol 12 (8) ◽

pp. 1817 ◽

Cited By ~ 1

Author(s):

Andrea A. Scamporrino ◽

Concetto Puglisi ◽

Angela Spina ◽

Maurizio Montaudo ◽

Daniela C. Zampino ◽

...

Keyword(s):

13C Nmr ◽

Microstructural Analysis ◽

Molar Fraction ◽

Polymer Chains ◽

Molar Ratio ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Cyclic Oligomers ◽

Ft Ir ◽

Tof Ms

New functionalized Poly(ether sulfone)s having different molar ratio (10, 20, 30, 50, 70, 100 mol%) of 4,4-bis phenoxy pentanoic acid unit (diphenolic acid; DPA) units were synthesized and characterized by (1H and 13C)-NMR, MALDI-TOF MS, FT-IR, DSC and DMA analyses. The microstructural analysis of the copolymers, obtained by 13C-NMR using an appropriate statistical model, shows a random distribution of copolymer sequences, as expected. The presence of different amount of DPA units along the polymer chains affects the chemical and physical properties of the copolymers. The Tg and the contact angle values decrease as the molar fraction of DPA units increases, whereas the hydrophilicity increases. NMR and MALDI-TOF MS analyses show that all polymer chains are almost terminated with hydroxyl and chlorine as end groups. The presence of cyclic oligomers was also observed by MALDI-TOF MS analysis.

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